Inhomogeneous distributions of ion channels, lipids, and carbohydrates in and on a cell membrane contribute to biophysical mechanisms of controlling single cell physiology. Fluorescence imaging, patch-based and whole-cell electrophysiology, differential scanning calorimetry, freeze fracture microscopy, and atomic force microscopy are some of the tools which have been used to provide information about cell membrane organization. These techniques are good at visualizing single proteins, measuring whole cell kinetics, or creating nanometer sized profiles of the membrane surface. However, an effective method for determining the local functional organization around a living cell membrane with temporal resolution has remained a challenge. Therefore, a need remains to develop methods to characterize the localized composition and dynamic attributes of living cells, particularly the cell membrane, and thereby increase our understanding of cellular physiology.